PAK Membrane Translocation and Phosphorylation Regulate Platelet Aggregation Downstream of Gi and G12/13 Pathways

Jianjun Zhang; Yan Zhang; Shuang Zheng; Yangyang Liu; Lin Chang; Guanxing Pan; Liang Hu; Si Zhang; Junling Liu; Soochong Kim; Jianzeng Dong; Zhongren Ding

doi:10.1055/s-0040-1714745

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2020; 120(11): 1536-1547
DOI: 10.1055/s-0040-1714745

Cellular Haemostasis and Platelets

PAK Membrane Translocation and Phosphorylation Regulate Platelet Aggregation Downstream of Gi and G_12/13 Pathways

Jianjun Zhang‡^*

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yan Zhang‡^*

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Shuang Zheng

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Yangyang Liu

²Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

,

Lin Chang

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Guanxing Pan

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Liang Hu

²Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

,

Si Zhang

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

,

Junling Liu

³Department of Biochemistry and Molecular Biology, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

,

Soochong Kim

⁴College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea

,

Jianzeng Dong

²Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

,

Zhongren Ding

¹Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

²Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

› Author Affiliations

Abstract

Platelet activation plays a pivotal role in physiological hemostasis and pathological thrombosis causing heart attack and stroke. Previous studies conclude that simultaneous activation of Gi and G_12/13 signaling pathways is sufficient to cause platelet aggregation. However, using Gq knockout mice and Gq-specific inhibitors, we here demonstrated that platelet aggregation downstream of coactivation of Gi and G_12/13 depends on agonist concentrations; coactivation of Gi and G_12/13 pathways only induces platelet aggregation under higher agonist concentrations. We confirmed Gi and G_12/13 pathway activation by showing cAMP (cyclic adenosine monophosphate) decrease and RhoA activation in platelets stimulated at both low and high agonist concentrations. Interestingly, we found that though Akt and PAK (p21-activated kinase) translocate to the platelet membrane upon both low and high agonist stimulation, membrane-translocated Akt and PAK only phosphorylate at high agonist concentrations, correlating well with platelet aggregation downstream of concomitant Gi and G_12/13 pathway activation. PAK inhibitor abolishes Akt phosphorylation, inhibits platelet aggregation in vitro and arterial thrombus formation in vivo. We propose that the PAK-PI3K/Akt pathway mediates platelet aggregation downstream of Gi and G_12/13, and PAK may represent a potential antiplatelet and antithrombotic target.

Keywords

platelets - thrombosis - Gi - G_12/13 - Akt - PAK

Full Text

References

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Supplementary Material

Supplementary Material